Clinical Laboratory Animal Medicine

Clinical Laboratory Animal

Medicine

An Introduction

Fifth Edition

Lesley A. Colby, DVM, MS, DACLAMMegan H. Nowland, DVM, DACLAMLucy H. Kennedy, DVM, DACLAM

This edition first published 2020© 2020 John Wiley & Sons, Inc.

Edition History4e 2013 John Wiley & Sons, Inc.; 3e 2007 Karen Hrapkiewicz and Leticia Medina; 2e 1998 © Iowa State University Press; 1e 1984 Iowa State University Press

All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or trans-mitted, in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, except as permitted by law. Advice on how to obtain permission to reuse material from this title is available at http://www.wiley.com/go/permissions.

The right of Lesley A. Colby, Megan H. Nowland, Lucy H. Kennedy to be identified as the authors of this work has been asserted in accordance with law.

Registered Office(s)John Wiley & Sons, Inc., 111 River Street, Hoboken, NJ 07030, USAJohn Wiley & Sons Ltd, The Atrium, Southern Gate, Chichester, West Sussex, PO19 8SQ, UK

Editorial Office111 River Street, Hoboken, NJ 07030, USA

For details of our global editorial offices, customer services, and more information about Wiley products visit us at www.wiley.com.

Wiley also publishes its books in a variety of electronic formats and by print‐on‐demand. Some content that appears in standard print versions of this book may not be available in other formats.

Limit of Liability/Disclaimer of WarrantyThe contents of this work are intended to further general scientific research, understanding, and discussion only and are not intended and should not be relied upon as recommending or promoting scientific method, diagnosis, or treatment by physicians for any particular patient. In view of ongoing research, equipment modifications, changes in governmental regulations, and the constant flow of information relating to the use of medicines, equipment, and devices, the reader is urged to review and evaluate the information provided in the package insert or instructions for each medicine, equipment, or device for, among other things, any changes in the instructions or indication of usage and for added warnings and precautions. While the pub-lisher and authors have used their best efforts in preparing this work, they make no representations or war-ranties with respect to the accuracy or completeness of the contents of this work and specifically disclaim all warranties, including without limitation any implied warranties of merchantability or fitness for a particular purpose. No warranty may be created or extended by sales representatives, written sales materials or promo-tional statements for this work. The fact that an organization, website, or product is referred to in this work as a citation and/or potential source of further information does not mean that the publisher and authors endorse the information or services the organization, website, or product may provide or recommendations it may make. This work is sold with the understanding that the publisher is not engaged in rendering professional services. The advice and strategies contained herein may not be suitable for your situation. You should consult with a specialist where appropriate. Further, readers should be aware that websites listed in this work may have changed or disappeared between when this work was written and when it is read. Neither the publisher nor authors shall be liable for any loss of profit or any other commercial damages, including but not limited to special, incidental, consequential, or other damages.

Library of Congress Cataloging‐in‐Publication Data

Names: Colby, Lesley, A., author. Title: Clinical laboratory animal medicine : an introduction / Lesley A. Colby, DVM, MS, DACLAM,

Megan H. Nowland, DVM, DACLAM, Lucy H. Kennedy, DVM, DACLAM. Description: Fifth edition. | Hoboken, NJ : Wiley-Blackwell, 2020. | Revision of: Clinical laboratory

animal medicine : an introduction / Karen Hrapkiewicz, Lesley Colby, Patricia Denison. Fourth edition. 2013. | Includes bibliographical references and index.

Identifiers: LCCN 2019030439 (print) | LCCN 2019030440 (ebook) | ISBN 9781119489566 (paperback) | ISBN 9781119489634 (adobe pdf) | ISBN 9781119489597 (epub)

Subjects: LCSH: Laboratory animals–Diseases. | Veterinary medicine. Classification: LCC SF996.5 .H65 2020 (print) | LCC SF996.5 (ebook) | DDC 636.088/5–dc23 LC record available at https://lccn.loc.gov/2019030439LC ebook record available at https://lccn.loc.gov/2019030440

Cover Design: WileyCover Images: Austin Thomason, Michigan Photography – University of Michigan; Amy Puffenberger, Animal Care & Use Program – University of Michigan

Set in 10/13pt Stempel Garamond by SPi Global, Pondicherry, India

10 9 8 7 6 5 4 3 2 1

I dedicate this book:

• To the memory of all the animals that have contributed both to my education and to the advancement of human and animal health. It has been a pleasure learning from you.

• To the animal technicians, veterinary technicians, and veterinarians who have devoted their lives to the care of laboratory animals and in support of animal welfare. It has been an honor working beside you.

• To my husband, Ben; our children, Nate and Tess; and our “special” family members (aka dogs), Troika, Ronal Danne, Tasha, and Bacca—for your incredible patience and support through all the nights, weekends, and holidays I spent studying or working over the years. Without you, I would not be who I am today.

LAC

To my husband, who supports me in all things whether large or unimportant, while reminding me not to take myself too seriously.

MHN

To Carolyn “Kit” Kestrel Leigh Kennedy, whose timely arrival made completing this book a difficult task! And to Scott and Mer, who made it possible anyways.

LHK

NOTE

The dosages given in this text are derived from published literature, but as few drugs are specifically licensed for use in the species described, the application is often extra‐label and may be empirical or based on clinical experience. The authors have made every attempt to verify all dosages and references; however, despite these efforts, errors in the original sources or in the preparation of this book may have occurred. Users of this text should exercise caution and evaluate all dosages prior to use to determine that they are reasonable.

vii

Contents

About the Authors xii

Preface xiii

About the Companion Website xiv

1 Introduction to Laboratory Animal Medicine 1Animals Used in Research, Teaching, and Testing 2Ethical Considerations 8Organizations 12Bibliography 17Further Reading 18Chapter 1 Review 19

2 Regulations, Policies, and Principles Governing the Care and Use of Laboratory Animals 22Animal Welfare Act and Regulations 22Public Health Service Policy on Humane Care And Use of Laboratory Animals 28Other Regulations, Policies, Guidance Documents, and Organizations 31References 36Further Reading 37Chapter 2 Review 37

3 Facility Design, Housing, Equipment, and Management 39Laboratory Animal Facility Design 39Common Facility Classifications 46Housing 50

viii Contents

Facility Equipment 59Management 66Bibliography 69Further Reading 69Chapter 3 Review 71

4 Mice 74Genetics 74Microbiologic Classifications 76Uses 77Behavior 77Anatomic and Physiologic Features 78Breeding and Reproduction 80Husbandry 81Techniques 86Special Techniques: Transgenic Production Technology 96Therapeutic Agents 101Introduction to Diseases of Mice 101Viral Diseases 107References 114Further Reading 119Chapter 4 Review 122

5 Rats 124Genetics 124Microbiologic Classifications 125Uses 126Behavior 126Anatomic and Physiologic Features 127Breeding and Reproduction 129Husbandry 130Techniques 133Therapeutic Agents 143Introduction to Diseases of Rats 144Bibliography 157Further Reading 162Chapter 5 Review 162

6 Gerbils 165Uses 165Behavior 166Anatomic and Physiologic Features 166Breeding and Reproduction 167Husbandry 168Techniques 170Therapeutic Agents 176Introduction to Diseases of Gerbils 176

Contents ix

Bibliography 181Further Reading 183Chapter 6 Review 184

7 Hamsters 185Uses 185Behavior 186Anatomic and Physiologic Features 187Breeding and Reproduction 189Husbandry 190Techniques 192Therapeutic Agents 198Introduction to Diseases of Hamsters 198Bibliography 207Further Reading 209Chapter 7 Review 210

8 Guinea Pigs 212Uses 212Behavior 213Anatomic and Physiologic Features 214Breeding and Reproduction 216Husbandry 218Techniques 219Therapeutic Agents 225Introduction to Diseases of Guinea Pigs 226Bibliography 238Further Reading 240Chapter 8 Review 242

9 Chinchillas 243Uses 243Behavior 244Anatomic and Physiologic Features 244Breeding and Reproduction 246Husbandry 247Techniques 249Therapeutic Agents 253Introduction to Diseases of Chinchillas 255Bibliography 260Further Reading 262Chapter 9 Review 262

10 Zebrafish 264Uses 264Behavior 265Anatomic and Physiologic Features 265

x Contents

Reproduction And Life Stages 266Husbandry 268Techniques 274Therapeutic Agents 278Introduction to Diseases of Zebrafish 278Bibliography 283Further Reading 284Chapter 10 Review 285

11 Rabbits 286Breeds 286Uses 287Behavior 288Anatomic and Physiologic Features 288Breeding and Reproduction 291Husbandry 293Techniques 297Therapeutic Agents 308Introduction to Diseases of Rabbits 309Bibliography 326Further Reading 329Chapter 11 Review 331

12 Ferrets 333Uses 333Behavior 334Anatomic and Physiologic Features 334Breeding and Reproduction 336Husbandry 337Techniques 339Therapeutic Agents 347Introduction to Diseases of Ferrets 347Bibliography 365Further Reading 369Chapter 12 Review 369

13 Primates 371Taxonomy 371Uses 375Behavior 376Anatomic and Physiologic Features 377Breeding and Reproduction 378Husbandry 380Techniques 383Therapeutic Agents 390Introduction to Diseases of Nonhuman Primates 390

Contents xi

Bibliography 410Further Reading 413Chapter 13 Review 416

14 Cattle, Sheep, Goats, and Pigs 417Uses 417Behavior 418Anatomic and Physiologic Features 419Breeding and Reproduction 419Husbandry 420Techniques 424Therapeutic Agents 428Introduction to Diseases of Agricultural Animals of Particular Importance to Research 428Bibliography 434Further Reading 435Chapter 14 Review 435

15 Research Variables, Biosecurity, and Colony Health Surveillance 437Research Variables 437Biosecurity and Exclusion of Contaminants 445Animal Colony Health Surveillance 448Bibliography 455Further Reading 455Chapter 15 Review 456

Appendix 1: Normal Values 459

Appendix 2: Comparative Biologic and Reproductive Values by Species 464

Answers to Review Questions 467

Index 474

xii

About the Authors

Lesley A. Colby, DVM, MS, DACLAM, is Associate Professor and Senior Director of Animal Resources and Operations in the Department of Comparative Medicine at the University of Washington (UW). She is Director of the UW’s BSL3/ABSL3 Facility and has particular interests in biocontainment, occupational health, and facility design and management. Dr. Colby is a Diplomate of the American College of Laboratory Animal Medicine.

Megan H. Nowland, DVM, DACLAM, is an Associate Professor in the Unit for Laboratory Animal Medicine at the University of Michigan in Ann Arbor, Michigan. There, she directs the Postdoctoral Training Program in Laboratory Animal Medicine, is the Associate Attending Veterinarian and the Assistant Director for Clinical Services.

Lucy H. Kennedy, DVM, DACLAM, is an Assistant Professor in the Unit for Laboratory Animal Medicine at the University of Michigan in Ann Arbor, Michigan. She is also the Managing Director for the Unit for Laboratory Animal Medicine’s Germ‐Free Mouse Facility and enjoys the challenges of gnotobiotic mouse research.

xiii

Preface

The purpose of this book is to provide basic information regarding the safe and responsible conduct of animal research as well as the unique anatomic and physio-logic characteristics, husbandry practices, and veterinary care of many of the animals frequently used in research: rodents, rabbits, ferrets, zebrafish, nonhuman primates, and agricultural animals. As the book name implies, we have designed the materials to be especially useful for individuals who are new to animal research as well as for those experienced in this field and expanding their knowledge of additional species. As a result, the book should be a useful resource for practicing veterinarians, veterinary students, veterinary technicians, research scientists, and others interested in learning about the field.

This, the fifth edition of the book, has been revised to include not only updated information but also new chapters on zebrafish and agricultural animals used in biomedical research. Significant changes have been made to expand and/or reorganize the non‐species chapters, to refine drug dosage tables to reflect the drugs most frequently utilized for each species, and to provide recommended reading sources for additional inquiry. As in previous editions of this book, study review questions are provided for each chapter and supplemental materials are provided in an accompanying website.

This book would not have been possible without the contributions of all previous edition authors: Donald Holmes, Karen Hrapkiewicz, Leticia Medina, and Patricia Denison. We thank you for providing such a strong foundation upon which to build. We also acknowledge and thank the many individuals and vendors who provided images for inclusion in the book and website. A very special thanks to our family, friends, and colleagues for their patience and support during the writing of this text. Lastly, our heartfelt thanks to all the animals that have contributed to the remarkable advancements in biomedical research throughout the ages, without whom many of the scientific breakthroughs we now take for granted would not have been possible.

Lesley A. ColbyMegan H. Nowland

Lucy H. Kennedy

xiv

About the Companion Website

This book is accompanied by a companion Website:

www.wiley.com/go/colby/clinical

The Website includes:

• Editable chapter review exercises and answers• Teaching PowerPoint presentations• Images in PowerPoint

Watch for throughout the book. These pinpoint materials that are also avail-able on the Website.

Clinical Laboratory Animal Medicine: An Introduction, Fifth Edition. Lesley A. Colby, Megan H. Nowland, and Lucy H. Kennedy. © 2020 John Wiley & Sons, Inc. Published 2020 by John Wiley & Sons, Inc.Companion website: www.wiley.com/go/colby/clinical

chap

ter

1

1

The health and welfare of animals used in biomedical research must be supported to be consistent with contemporary ethical standards and to help ensure the scientific validity of research results. Providing this support requires individuals with expertise in many fields including basic and applied sciences, bioethics, regulatory oversight, experimental design, and laboratory animal science. Laboratory animal science is defined by the US National Library of Medicine as “[t]he science and technology dealing with the procurement, breeding, care, health, and selection of animals used in biomedical research and testing” (Box 1.1). It includes husbandry, nutrition, behavior, health care, production, and management of laboratory animals.

Laboratory animal medicine is a specialized field within laboratory animal science and a recognized specialty within veterinary medicine. At its core, laboratory animal medicine encompasses the diagnosis, treatment, and prevention of diseases in animals used in research, teaching, and testing. It emphasizes methods to prevent and mini-mize pain, discomfort, and distress in research animals; facilitates acquisition of bio-logically meaningful results; and minimizes experimental variability. The field has progressively grown and evolved in response to scientific and medical advances, shifts in the regulatory environment, and the ever‐changing focus of scientific inquiry.

Diverse groups of individuals play important roles within laboratory animal med-icine. Veterinarians have a variety of responsibilities within an animal care and use program that may include provision of veterinary care, management of animal care

Introduction to Laboratory Animal Medicine

1

2 Clinical Laboratory Animal Medicinech

apte

r 1

and use facilities, education of individuals who care for and use laboratory animals, assisting biomedical scientists in the selection of and humane use of animals, obtaining and interpreting biologically relevant data, and assuring compliance with regulations and policies that affect research animals. Veterinary technicians work under the super-vision of a veterinarian, assisting them in carrying out these responsibilities. They often provide technical support in disease detection, including oversight of colony health monitoring programs, treatment of ill animals, blood sampling, and necropsy and tissue collection. When engaged in research or drug study positions at a pharmaceutical firm or university, they administer test products and collect data. This type of employment often requires the veterinary technician to be credentialed and have a bachelor’s degree. Credentialed veterinary technicians, with sufficient education, training, and experience, and who have completed testing requirements can apply to join the Academy of Laboratory Animal Veterinary Technicians and Nurses, a specialty organization within the National Association of Veterinary Technicians in America (NAVTA). Veterinary technicians may also work in research compliance or supervise other animal facility staff such as assistant laboratory animal technicians, animal caretakers, and cagewash per-sonnel. It should also be noted that individuals without formal veterinary training make significant contributions in support of laboratory animal medicine. For example, animal caretakers who closely observe and handle animals on a daily basis can be instrumental in detecting behavioral changes and identifying early signs of illness so that animals can be promptly assessed by veterinary personnel.

ANIMALS USED IN RESEARCH, TEACHING, AND TESTING

Biomedical ResearchRemarkable advances have been made in medicine and science over the past century, such as the characterization of complex host–pathogen interactions and immune system functions, development of vaccines for polio and hepatitis B, creation of antibiotics and antivirals for infectious diseases, procedures for organ transplantation and open heart surgery, and development of drugs for chronic disorders such as diabetes and high blood pressure. Animals played a major role in each of these advances (Table 1.1). New treatment modalities for cancer, less invasive surgical approaches, and the development of equipment such as the laser and endoscopic instruments would not have been pos-sible without the use of animals. Often, advances made in human health are also applied to the benefit of companion animals (Figure 1.1). For instance, most cancer treatments and many advanced surgical techniques and imaging modalities developed for use in humans are now routinely available to veterinary practices.

Box 1.1

The US National Library of Medicine defines laboratory animal science as “[t]he science and technology dealing with the procurement, breeding, care, health, and selection of animals used in biomedical research and testing.”

Introduction to Laboratory Animal Medicine 3

chap

ter

1Table 1.1. Animal roles in medical discoveries and advancements

Year* Scientist(s) Animal(s) Used Contribution

1901 von Behring Guinea pig Development of diphtheria antiserum

1904 Pavlov Dog Animal responses to various stimuli

1923 Banting, Macleod Dog, rabbit, fish Discovery of insulin and mechanism of

diabetes

1924 Einthoven Dog Mechanism of the electrocardiogram

1945 Fleming, Chain, Florey Mouse Discovery of penicillin and its curative

effect in various infectious diseases

1954 Enders, Weller, Robbins Monkey, mouse Culture of poliovirus that led to

development of vaccine

1964 Block, Lynen Rat Regulation of cholesterol and fatty acid

metabolism

1966 Rous Rat, rabbit, hen Discoveries concerning hormonal

treatment of prostatic cancer

1970 Katz, von Euler, Axelrod Cat, rat Mechanism of storage and release of

nerve transmitters

1979 Cormack, Hounsfield Pig Development of computer‐assisted

tomography (CAT scan)

1984 Milstein, Koehler, Jerne Mouse Techniques of monoclonal antibody

formation

1990 Murray, Thomas Dog Organ transplant techniques

1997 Prusiner Mouse, hamster Discovery of prions, a new biological

principle of infection

2003 Lauterbur, Mansfield Clam, mouse, dog, rat,

chimpanzee, pig,

rabbit, frog

Discoveries concerning magnetic

resonance imaging

2008 Barre‐Sinoussi, Montagnier Monkey, chimpanzee,

mouse

Discovery of human immunodeficiency

virus

2008 zur Hausen Hamster, mouse, cow Discovery of papilloma viruses causing

cervical cancer

2011 Hoffman, Beutler Fruit fly, mouse Discoveries concerning the activation of

innate immunity

2011 Steinman Mouse Discovery of the dendritic cell and its

role in adaptive immunity

2012 Gurdon, Yamanaka Frog, mice Discovery that mature cells can be

reprogrammed to become pluripotent

2013 Rothman, Schekman,

Sudhop

Mouse, hamster Discovery of how cells organize

movement of materials into and out

of cells

2014 O’Keefe, Britt, Moser Rat Discovery of cells that constitute the

brain’s “inner GPS” positioning

system

(Continued )

4 Clinical Laboratory Animal Medicinech

apte

r 1

Significant advances have been made in the development and use of ex vivo (“out of the living”) experimental methods which do not require the use of animals or animal‐derived products. These experimental methods should be used in place of in vivo (“in life”) experimental methods whenever possible, but only when resultant experimental findings are truly representative and predictive of the system(s) they are intended to model (Box 1.2). Unfortunately, most ex vivo testing systems cannot generate sufficiently comprehensive and accurate data representative of an intricate, living being. As a result, use of in vivo experimental methods is still required until more refined alternatives are developed and validated. In the interim, use of ex vivo experimental methods can be effective and valuable in refining and reducing animal use for some areas of study such as early identification of toxic or ineffective experi-mental compounds and modeling compound–receptor interactions.

TeachingAnimals play a valuable role in education, starting from preschool and continuing to the college and graduate levels. Although computer modeling and videos can replace select learning experiences, some personal learning styles and educational objectives are best suited to hands‐on learning. Through interactions with animals, children can learn how to care for another living being. They also learn lessons in responsibility and respect. At the middle and high school levels, animal tissues may be used for hands‐on experience with dissection and now‐common laboratory methods such as immunoassays and molecular diagnostics. These experiences often reveal the amazing world of biology and science to young people as they learn about the complex and specialized processes that form the basis of biological functions. In college, animals

Table 1.1. (Continued )

Year* Scientist(s) Animal(s) Used Contribution

2015 Campbell, Omura, Tu Mouse, dog, sheep,

cattle, chicken, monkey

Discoveries contributing to development

of novel therapies for roundworm

and for malarial infections

2016 Ohsumi Mouse Discovery of mechanisms for cellular

autophagy

2017 Rosbash, Hall, Young Fruit flies Discovery of molecular mechanisms

controlling the circadian rhythm

Sources: National Association of Biomedical Research (https://www.nabr.org/biomedical‐research/

medical‐progress), Foundation for Biomedical Research (https://fbresearch.org/medical‐advances/nobel‐

prizes), and Nobel Prize (www.nobelprize.org).

* Year of occurrence or award recognition.

Box 1.2

Ex vivo experimental methods should be used in place of in vivo experimental methods whenever possible.

Introduction to Laboratory Animal Medicine 5

chap

ter

1are used in a variety of professional and graduate‐level courses in medical and health‐related fields. Surgery courses provide young veterinary surgeons a chance to hone their skills before performing them on client‐owned animals. Physicians use animals to practice robotic, endoscopic, and laser surgery prior to performing them in people. Animals are used in training courses for medical personnel so they may acquire and advance their skills in emergency and critical care environments. Clinical technique courses provide veterinary students and veterinary technician students opportunities to develop the diverse range of skills necessary for the safe and humane care of their future animal patients, such as animal handling and performance of physical exams, injections, and catheterizations. It is now common for animals used for educational purposes to either be temporarily housed in a research/teaching envi-ronment and then adopted into loving homes or be pets brought in by their owners to assist with training.

Product Safety TestingSeveral decades ago, consumers were subjected to publicly marketed drugs and cosmetics that were not adequately tested to assess human safety. Examples included early treatments for syphilis containing mercury and arsenic, an eyelash dye that caused blindness in numerous individuals, and an elixir marketed for use by children and that caused the death of over 100 people. These and other similar events led to the passage of the Food, Drug, and Cosmetic Act (FD&C) in 1938. Broadly speaking, the FD&C was created to safeguard and protect consumer health and safety from the sale of dangerous products. The Act is enforced pri-marily by the Food and Drug Administration (FDA) which requires animal testing of products when a scientifically valid, alternative testing method is not available. When animal testing is required to obtain FDA approval, the FDA requires prod-uct manufacturers and sponsors to conduct the studies in accordance with the Good Laboratory Practice for Nonclinical Laboratory Studies (21 CFR Part 58). Recently, many manufacturers have marketed cosmetic products as “cruelty‐free.” This unregulated labeling or advertising practice can be deceiving to consumers who may incorrectly assume that neither the product nor any product components were tested in animals. However, it is more likely that animal safety testing had been performed on individual product components (but not necessarily the final product) or that testing had been contracted by the manufacturer to be performed by an external testing group. As a consequence, consumers must be savvy in their interpretation of unregulated product labeling.

Animal Usage StatisticsAccording to the US Department of Agriculture (USDA) Animal Report Animal Usage by Fiscal Year, 792,168 animals whose use is regulated by the Animal Welfare Act were used for research, teaching, and product safety testing in the United States in 2017. It should be noted that this figure does not include the annual usage of mice, rats, birds, or fish as the USDA is not charged with regulatory oversight of these species as required by Animal Welfare Act. Rather, use of these species is regulated by other entities and the precise numbers of their use is unknown. However, it is esti-mated that up to 26 million animals of these species are used annually. Mice and rats

6 Clinical Laboratory Animal Medicinech

apte

r 1

account for greater than 95% of all animals used while the number of dogs, cats, and nonhuman primates combined account for less than 1% of the animals used.

With the exception of the increased use of zebrafish, the use of nonrodent animals has been declining over the past three decades primarily due to use of more refined experimental systems and an intense effort by the biomedical research community to

Fig 1.1. Animal research saves animals too. (Source: Foundation for Biomedical Research.)

Introduction to Laboratory Animal Medicine 7

chap

ter

1

decrease animal use (Box 1.3). The number of dogs used in research currently is less than one‐third of its numbers in the late 1970s. The number of nonhuman primates used over the past decade has risen slightly, in part due to increasing emphasis of research into human brain function and neurodegenerative diseases such as Alzheimer’s. Across all species, the majority of animals used in biomedical research are bred specifically for that purpose.

To put the numbers in perspective, approximately 12–27 million animals are used in research in the United States. All but approximately 1 million of these are mice, rats, birds, or fish. According to Speaking of Research (www.speakingofresearch.com), “we consume over 1800 times the number of pigs than the number [of pigs] used in research” and we consume over 340 times more chickens than the total number of animals used in biomedical research.

Funding SourcesIn the United States, the National Institutes of Health (NIH) and the National Science Foundation (NSF) are the primary public granting agencies for biomedical research. The NIH, a branch of the Public Health Service (PHS), provides competitive federal grants for investigators interested in the health‐related advancement of humans and animals. The NSF encourages basic research in behavior, mathematics, physics, medicine, biology, and other sciences. In addition to the NIH and NSF, funding is available from universities and colleges, state governments, industry, and private foundations. Acquiring funds to conduct research is difficult as competition for grant money is high, with only 10%–20% of submitted proposals receiving funding. Typically, a grant provides money for the primary scientist’s and research team’s salaries, supplies, equipment, and purchase and care of animals for a 3‐year period. The primary scientist, or principal investigator (PI), is responsible for planning and coordinating all phases of the research study, including tabulating data, reporting findings to the funding agency, and publication of results. When a study yields valuable results, the funding agency may renew the grant for an additional period of time.

Regulatory Oversight and AccreditationMultiple levels of regulation (e.g., federal, state, and local) function to provide oversight of animal research including mandating standards for animal care and use. In addition, many institutions choose to participate in voluntary assessment and accreditation programs which recognize institutions that have exceeded the minimum standards required by law and have achieved excellence in animal care and use. Chapter  2 provides additional information regarding the oversight provided by governmental and voluntary organizations.

Box 1.3

With the exception of the increased use of zebrafish, the use of nonrodent animals has been declining over the past three decades.

8 Clinical Laboratory Animal Medicinech

apte

r 1 Institutional Animal Care and Use Committee

Prior to the use of animals in research, teaching, or testing, a protocol must be submitted to and approved by the institution’s Institutional Animal Care and Use Committee (IACUC) (Box 1.4). The protocol is a detailed, written description of the proposed animal care and use. It justifies the use of vertebrate animals to accom-plish the study’s aims, details the procedures that will be performed on the animals, and describes how the animals will be housed and cared for throughout the project. Additionally, the PI must give several assurances, including that the study does not unnecessarily duplicate previous studies, that the staff working with the animals have adequate training to accomplish the study tasks in a humane manner, that alternatives to animal use have been carefully considered, and that any activities that may induce animal pain or distress are scientifically necessary. Animal use protocols are usually approved for 3 years, may undergo an annual review by the IACUC, and must be resubmitted for full, de novo (anew) review every 3 years. IACUCs must formally review and approve all changes to approved protocols prior to their implementation. Moreover, IACUCs are required, at a minimum frequency (usually twice yearly), to review their institution’s established program of animal care and use and physically inspect facilities where animals are housed or manipulated. Additional information about IACUCs can be found at www.iacuc.org.

ETHICAL CONSIDERATIONS

The 3Rs: Replacement, Refinement, and ReductionTwo English scientists, Russell and Burch, coined the term “the 3Rs.” In 1959, they examined the ethical aspects and “the development and progress of humane tech-niques in the laboratory” (Russel and Burch, 1959). The 3Rs represent three ethical tenets of responsible animal use: replacement, refinement, and reduction (Box 1.5). Research institutions and regulatory authorities continually strive to apply the prin-ciples of the 3Rs to ensure animals are used in an ethical manner. There is an ethical imperative that scientists use animals only when they have provided assurance to the

Box 1.5

Russel and Burch’s 3Rs represent three ethical tenets of responsible animal use: replacement, refinement, and reduction.

Box 1.4

Prior to the use of animals in research, teaching, or testing, a protocol must be submitted to and approved by the institution’s Institutional Animal Care and Use Committee (IACUC).

Introduction to Laboratory Animal Medicine 9

chap

ter

1IACUC that no nonanimal methods will allow them to achieve their scientific aim. This search for alternatives is mandated for species covered by the Animal Welfare Act (AWA; see APHIS, 2017). For species not covered by the AWA, both the Public Health Service Policy and the Guide for the Care and Use of Research Animals (ILAR, 2011; the Guide) refer to the “US Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training” (OLAW, 1985), which includes language about alternatives to animal use (see Chapter 2, Table 2.1). The US Government Principles also mandate using the minimum number of animals necessary to obtain valid results. This is synonymous with reduction, one of the 3Rs.

Replacement refers to replacing animals with a nonanimal alternative, such as in vitro (“in glass,” outside of the body) screens with cell culture or computer (in silica) modeling, or by using the least sentient animal (e.g., rat in place of dog; fish in place of mouse) that will enable collection of meaningful and valid data. Continued advances in the sciences and testing methods have helped to spur development of animal testing alternatives. Any alternative test, however, must be validated before it can be used to replace a test currently using animals. The development and use of genetically special-ized animals, such as nude and transgenic mice, has made it possible to reduce the number of other species such as dogs and cats. Environmental toxicity studies often use zebrafish rather than mice or other mammals. Alternate tests for ophthalmic safety testing have been developed using tissues obtained from slaughterhouses as well as specially designed cell and tissue culture systems. In addition, the limulus ame-bocyte lysate (LAL) assay has largely replaced the rabbit pyrogen test for detecting pyrogens, such as endotoxin, in injectable substances.

Refinement refers to methods that incorporate modification of a procedure to lessen animal pain and distress or enhance animal well‐being. Use of less invasive pro-cedures, provision of pain relief, provision of environmental enrichment, and decreased restraint time are examples of refinements. For example, through advanced imaging techniques, such as magnetic resonance imaging (MRI), researchers can now view structures and observe anatomic functions that once could only be accomplished during surgery or at necropsy. Investigators must constantly review the way animal studies are conducted to ensure that the methods used are the most humane and refined to minimize pain and distress. In addition, investigators work closely with laboratory animal veterinarians and the IACUC to assure that humane experimental endpoints are in place to minimize pain and distress to the greatest degree possible. The IACUC often collaborates with investigators and veterinary personnel to develop humane endpoint guidelines that help determine when an animal should be eutha-nized or removed from a study. Examples of humane experimental endpoints include a defined percentage of weight loss, tumor size, presence of labored breathing, or an inability to ambulate. There is a delicate balance between collecting the necessary scientific data from a study and ensuring that animal welfare is preserved. For example, it can be difficult to identify the point at which an animal should be removed from study or euthanized before it becomes significantly ill. When appropriate, the least invasive experimental methods should be used, and anesthesia or analgesia be admin-istered to eliminate unnecessary pain and distress.

Reduction refers to using the minimal number of animals in a study while remain-ing consistent with sound scientific and statistical standards. Investigators must

10 Clinical Laboratory Animal Medicinech

apte

r 1 constantly strive to find ways to reduce animal numbers. Using a combination of

computer‐based simulators in conjunction with animal subjects, employing better statistical methods, or using one control group with multiple study groups are examples of methods used to reduce animal numbers. The number of animals used in product safety testing has been significantly reduced through validation of alternative testing methods. Experiments can be designed using multiple sections with the results derived from earlier sections used to refine the number of animals or experimental groups used in later sections. For example, a “staircase design” is often used in acute toxicology testing. This method involves administration of a limited number of drug dosages (high and low) to then determine a more precise dose range for further testing. Used with sophisticated computer‐assisted computa-tional methods, the staircase design can determine a point estimate of the lethal dose, approximate confidence intervals, and determine toxic signs for the substance tested, yet use fewer animals.

Overall, the research community must continually challenge itself to consider whether the animal research being performed is ethical and justifiable. The principles underlying the “3Rs” should be observed so that animal use in biomedical research is minimized while at the same time, data obtained from animal research is optimized. Only in that way will we be assured of continued public support for the animal research that benefits so much of society, including the health and welfare of nonhuman animals!

Animal Rights and Animal WelfareThe terms “animal rights” and “animal welfare” are not synonymous. Animal rights represents a philosophical belief that gives animals the same equality and protection as humans (Box 1.6). According to this philosophy, a field mouse has the same right to life as a human. Animal rights purports that animals should not be regarded as property. No matter how humane, animal use is viewed as exploitation and should be banned. This includes keeping dogs and cats as pets; displaying animals in zoos and aquariums; using chickens, cattle, or swine for food; and using animals in research, teaching, and testing. Furthermore, adherence to this philosophy prohibits one’s use of medications including vaccines and medical treatments that were developed through animal research.

Animal welfare represents a philosophical belief that it is morally acceptable for humans to use animals provided they are treated humanely and their physical and psychological well‐being is met (Box 1.7). This philosophy is based on a belief that animals can contribute to human welfare. Animals provide companionship, entertain-ment, labor, food, fiber, and advancement of knowledge when used in research and teaching. When animals are used, it is paramount that responsible practices of animal

Box 1.6

Animal rights represents a philosophical belief that gives animals the same equality and protection as humans.

Introduction to Laboratory Animal Medicine 11

chap

ter

1

welfare are adhered to, including provision of appropriate housing, handling, management, disease prevention and treatment, and, when necessary, euthanasia.

Sources of Animals Used in Biomedical ResearchAnimals used in research may be obtained from a variety of sources. The sale of AWA‐covered species (e.g., dogs and cats) to research facilities is regulated by the  USDA who licenses Class A and Class B animal dealers. USDA‐licensed Class A dealers supply “purpose bred” animals, animals bred and raised specifically for use in research. Purpose‐bred animals are of genetically similar back-grounds,  often with defined pedigrees, and have well‐documented health his-tories. To help ensure that these animals are accustomed to the research environment and are easy to handle, many vendors have instituted robust animal handling and socialization programs as components of their animal care programs.

A small number of animals used in research are obtained from USDA Class B dealers, who acquire animals from “random sources” such as individual owners, hobby breeders, and pounds and shelters. Although Class B dealers are subject to federal legislation under the Animal Welfare Act and are licensed by the USDA, public concern regarding acquisition of dogs and cats from Class B dealers led to the decision by NIH to discontinue funding of experiments using random source dogs and cats. Institutions do occasionally elect to obtain animals directly from random sources when purpose‐bred animals do not possess the characteristics necessary for study, such as advanced age or preexisting health conditions. Acquisition of these animals is tightly regulated.

Nonhuman Primate UseNonhuman primates are human’s closest genetic relatives. Due to this and their associated high level of sentience, their use in research should be reserved only for when another animal model cannot be used. Nonhuman primates account for less than 1% of the USDA‐regulated animals used in the United States. The vast majority of nonhuman primates used are rhesus and cynomolgus macaques. Although the use of chimpan-zees was invaluable in advancing human health, including for the development of vaccines for polio and hepatitis B, the NIH no longer supports the use of chimps in research and all use of chimps in research has been significantly restricted and effec-tively eliminated. Chimps previously used in research have been retired to designated sanctuaries or have been retired “in place” when it was reasonably expected that they would experience harm if relocated.

Box 1.7

Animal welfare represents a philosophical belief that it is morally acceptable for humans to use animals provided they are treated humanely and their physical and psychological well‐being is met.

12 Clinical Laboratory Animal Medicinech

apte

r 1 ORGANIZATIONS

The need for more systematic and specialized information on laboratory animal husbandry, medical care, and management of animal facilities and the desire to foster collaborative environments led to the development of several organizations that support the laboratory animal science community. The following is an intro-duction to some of the most important organizations and a brief description of their purpose.

American Association for Laboratory Animal ScienceIn 1950, the Animal Care Panel (ACP), a national professional organization dedicated to the care, production, and study of laboratory animals, was established. In 1967, the ACP became the American Association for Laboratory Animal Science (AALAS) whose mission is to advance “responsible laboratory animal care and use to benefit people and animals.” AALAS is a nonprofit, professional association that serves as the principal means of communication between individuals and organizations within the field of laboratory animal science. AALAS currently has over 14,500 individual and institutional members and more than 43 local branches. AALAS produces two scientific journals, Comparative Medicine and Journal of the American Association for Laboratory Animal Science, and several technician‐targeted publications including the quarterly magazine, Laboratory Animal Science Professional. AALAS also certifies trained technicians; promotes education through publications; supports the AALAS Learning Library, an extensive Web‐based continuing education site; and hosts an annual national meeting. Scientists, veterinarians, technicians, managers, and suppliers share information through presentations, discussions, and exhibits at the annual meeting. Recently, AALAS has increased its role in public outreach and promoting the benefits of biomedical research. For further information, visit www.aalas.org.

AALAS administers the AALAS Technician Certification Program through which technicians receive certification at one of three levels: Assistant Laboratory Animal Technician (ALAT), Laboratory Animal Technician (LAT), and Laboratory Animal Technologist (LATG) (Figure 1.2). The minimum qualifications required to take each certification exam are listed in Figure 1.3. The duties of assistant laboratory animal technicians are primarily related to animal care and facility sanitation. Laboratory animal technicians are expected to have increased diagnostic and technical skills and research responsibilities. Laboratory animal technologists are frequently involved in

Fig 1.2. AALAS technician certification level logos. (Source: AALAS.)

Introduction to Laboratory Animal Medicine 13

chap

ter

1

supervisory capacities and conducting portions of the research study. The achievement of certification at any level denotes an individual dedicated to the pursuit of a higher standard of technical skill and knowledge. Many institutions now require or prefer AALAS certification as a prerequisite for obtaining jobs in their animal facility. Alternatively, many institutions encourage employees to pursue certification as a means of advancing their careers and offer classes as part of their training programs. AALAS offers training manuals for each of the three levels and suggests other mate-rials appropriate for examination preparation. Employers often provide employees with financial support for the examinations and frequently reward the achievement of certification with a specific increase in salary.

AALAS also sponsors the Certified Manager of Animal Resources (CMAR) program. The CMAR designation is a sign of professionalism in the field of animal resources management. Certification requires successful completion of a series of general business management exams offered through the Institute for Certified Professional Managers (ICPM) or through AALAS as well as completion of the more specialized Animal Resources Exam offered by AALAS. Educational materials for the exams are available from ICPM and from AALAS, including AALAS’ Management Training Manual. The minimum eligibility requirements for CMAR designation are

Eligibility requirementsBelow are the minimum eligibility requirements for each exam.To be eligible for the exam you wish to take, you must meetone of the combinations of education and work experience.

Education level

Currentcert.level

ALATExam

LATExam

LATGExam

LAT

ALAT

ALAT

2

1

0.5

3

5

4

3

2

1

0.5*

0.5*

* Work experience must be acquired after attaining the specified certification.** Option for those without documentation of education level.

2**

HS/GED

orhigher

AA/ASor

higher

BA/BSor

higher

Lab animalwork

experience(years)

Fig 1.3. Minimum eligibility requirements for AALAS technician certification. (Source: AALAS.)

14 Clinical Laboratory Animal Medicinech

apte

r 1

listed in Table 1.2. CMAR recipients must fulfill continuing education requirements to maintain their designation.

Laboratory Animal Management AssociationThe Laboratory Animal Management Association (LAMA) was established in 1984 with the mission of “enhancing the quality of management and care of laboratory ani-mals throughout the world” by “promot[ing] education, knowledge exchange and professional development” of facility managers and supervisors (www.lama‐online.org). The organization publishes a quarterly journal, LAMA Review, sponsors training sessions, and hosts an annual educational meeting. For further information, visit www.lama‐online.org.

American Society of Laboratory Animal PractitionersIn August 1966, the Laboratory Animal Welfare Act became law and mandated that “adequate veterinary care” be provided to select laboratory animals. The American Society of Laboratory Animal Practitioners (ASLAP) was founded later that same year, partially in response to the Act’s passage. ASLAP is a professional organization through which veterinarians engaged or interested in the practice of laboratory animal medicine can freely exchange ideas, experiences, and knowledge. In 1967, ASLAP was officially recognized as an ancillary organization of the American Veterinary Medical Association (AVMA) and in 1986, ASLAP became an affiliate of AALAS. Both veter-inarians and veterinary students make up the membership of ASLAP. According to its website, the objectives of ASLAP are to (1) “provide a mechanism for the exchange of scientific and technical information among veterinarians engaged in laboratory animal practice,” (2) “actively encourage its members to provide training for veterinarians in the field of laboratory animal practice at both the pre and postdoctoral levels and lend their expertise to institutions conducting laboratory animal medicine programs,” (3) “encourage the development and dissemination of knowledge in areas related to lab-oratory animal practice,” and (4) “act as a spokesperson for laboratory animal practi-tioners within the AVMA House of Delegates and to work with other organizations involved in the care and use of laboratory animals in representing our common inter-ests and concerns to the scientific community and the public at large.” For further information, visit www.aslap.org.

Table 1.2. Eligibility requirements for CMAR designation

Education Level Total Work Experience Total Management Experience

BA/BS 5 years 3 years

AA/AS 8 years 3 years

HS/GED 10 years 3 years

Source: AALAS.

Note: Candidates meeting these requirements who pass the Animal Resources Exam and the Certified

Manager (CM) exams will achieve the status of a Certified Manager of Animal Resources and will be able

to use the CMAR acronym after their names.